A closer look at Aspergillus: online monitoring via scattered light enables reproducible phenotyping
Abstract Background Filamentously growing microorganisms offer unique advantages for biotechnological processes, such as extraordinary secretion capacities, going along with multiple obstacles due to their complex morphology. However, limited experimental throughput in bioprocess development still h...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2019-08-01
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| Series: | Fungal Biology and Biotechnology |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s40694-019-0073-x |
| _version_ | 1818684547199401984 |
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| author | Roman P. Jansen Carina Beuck Matthias Moch Bianca Klein Kira Küsters Holger Morschett Wolfgang Wiechert Marco Oldiges |
| author_facet | Roman P. Jansen Carina Beuck Matthias Moch Bianca Klein Kira Küsters Holger Morschett Wolfgang Wiechert Marco Oldiges |
| author_sort | Roman P. Jansen |
| collection | DOAJ |
| description | Abstract Background Filamentously growing microorganisms offer unique advantages for biotechnological processes, such as extraordinary secretion capacities, going along with multiple obstacles due to their complex morphology. However, limited experimental throughput in bioprocess development still hampers taking advantage of their full potential. Miniaturization and automation are powerful tools to accelerate bioprocess development, but so far the application of such technologies has mainly been focused on non-filamentous systems. During cultivation, filamentous fungi can undergo remarkable morphological changes, creating challenging cultivation conditions. Depending on the process and product, only one specific state of morphology may be advantageous to achieve e.g. optimal productivity or yield. Different approaches to control morphology have been investigated, such as microparticle enhanced cultivation. However, the addition of solid microparticles impedes the optical measurements typically used by microbioreactor systems and thus alternatives are needed. Results Aspergillus giganteus IfGB 0902 was used as a model system to develop a time-efficient and robust workflow allowing microscale cultivation with increased throughput. The effect of microtiter plate geometry, shaking frequency and medium additives (talc and calcium chloride) on homogeneity of culture morphology as well as reproducibility were analyzed via online biomass measurement, microscopic imaging and cell dry weight. While addition of talc severely affected online measurements, 2% (w v−1) calcium chloride was successfully applied to obtain a highly reproducible growth behavior with homogenous morphology. Furthermore, the influence of small amounts of complex components was investigated for the applied model strain. By correlation to cell dry weight, it could be shown that optical measurements are a suitable signal for biomass concentration. However, each correlation is only applicable for a specific set of cultivation parameters. These optimized conditions were used in micro as well as lab-scale bioreactor cultivation in order to verify the reproducibility and scalability of the setup. Conclusion A robust workflow for A. giganteus was developed, allowing for reproducible microscale cultivation with online monitoring, where calcium chloride is an useful alternative to microparticle enhanced cultivation in order to control the morphology. Independent of the cultivation volume, comparable phenotypes were observed in microtiter plates and in lab-scale bioreactor. |
| first_indexed | 2024-12-17T10:52:22Z |
| format | Article |
| id | doaj.art-636399c95b8649c0b8cb765b3c9a99d7 |
| institution | Directory Open Access Journal |
| issn | 2054-3085 |
| language | English |
| last_indexed | 2024-12-17T10:52:22Z |
| publishDate | 2019-08-01 |
| publisher | BMC |
| record_format | Article |
| series | Fungal Biology and Biotechnology |
| spelling | doaj.art-636399c95b8649c0b8cb765b3c9a99d72022-12-21T21:51:56ZengBMCFungal Biology and Biotechnology2054-30852019-08-016111110.1186/s40694-019-0073-xA closer look at Aspergillus: online monitoring via scattered light enables reproducible phenotypingRoman P. Jansen0Carina Beuck1Matthias Moch2Bianca Klein3Kira Küsters4Holger Morschett5Wolfgang Wiechert6Marco Oldiges7Forschungszentrum Jülich, Institute of Bio- and Geosciences-Biotechnology (IBG-1)Forschungszentrum Jülich, Institute of Bio- and Geosciences-Biotechnology (IBG-1)Forschungszentrum Jülich, Institute of Bio- and Geosciences-Biotechnology (IBG-1)Forschungszentrum Jülich, Institute of Bio- and Geosciences-Biotechnology (IBG-1)Forschungszentrum Jülich, Institute of Bio- and Geosciences-Biotechnology (IBG-1)Forschungszentrum Jülich, Institute of Bio- and Geosciences-Biotechnology (IBG-1)Forschungszentrum Jülich, Institute of Bio- and Geosciences-Biotechnology (IBG-1)Forschungszentrum Jülich, Institute of Bio- and Geosciences-Biotechnology (IBG-1)Abstract Background Filamentously growing microorganisms offer unique advantages for biotechnological processes, such as extraordinary secretion capacities, going along with multiple obstacles due to their complex morphology. However, limited experimental throughput in bioprocess development still hampers taking advantage of their full potential. Miniaturization and automation are powerful tools to accelerate bioprocess development, but so far the application of such technologies has mainly been focused on non-filamentous systems. During cultivation, filamentous fungi can undergo remarkable morphological changes, creating challenging cultivation conditions. Depending on the process and product, only one specific state of morphology may be advantageous to achieve e.g. optimal productivity or yield. Different approaches to control morphology have been investigated, such as microparticle enhanced cultivation. However, the addition of solid microparticles impedes the optical measurements typically used by microbioreactor systems and thus alternatives are needed. Results Aspergillus giganteus IfGB 0902 was used as a model system to develop a time-efficient and robust workflow allowing microscale cultivation with increased throughput. The effect of microtiter plate geometry, shaking frequency and medium additives (talc and calcium chloride) on homogeneity of culture morphology as well as reproducibility were analyzed via online biomass measurement, microscopic imaging and cell dry weight. While addition of talc severely affected online measurements, 2% (w v−1) calcium chloride was successfully applied to obtain a highly reproducible growth behavior with homogenous morphology. Furthermore, the influence of small amounts of complex components was investigated for the applied model strain. By correlation to cell dry weight, it could be shown that optical measurements are a suitable signal for biomass concentration. However, each correlation is only applicable for a specific set of cultivation parameters. These optimized conditions were used in micro as well as lab-scale bioreactor cultivation in order to verify the reproducibility and scalability of the setup. Conclusion A robust workflow for A. giganteus was developed, allowing for reproducible microscale cultivation with online monitoring, where calcium chloride is an useful alternative to microparticle enhanced cultivation in order to control the morphology. Independent of the cultivation volume, comparable phenotypes were observed in microtiter plates and in lab-scale bioreactor.http://link.springer.com/article/10.1186/s40694-019-0073-xAspergillus giganteusMicrotiter plateMiniaturized cultivationAntifungal proteinHigh throughput bioprocess developmentMorphology engineering |
| spellingShingle | Roman P. Jansen Carina Beuck Matthias Moch Bianca Klein Kira Küsters Holger Morschett Wolfgang Wiechert Marco Oldiges A closer look at Aspergillus: online monitoring via scattered light enables reproducible phenotyping Fungal Biology and Biotechnology Aspergillus giganteus Microtiter plate Miniaturized cultivation Antifungal protein High throughput bioprocess development Morphology engineering |
| title | A closer look at Aspergillus: online monitoring via scattered light enables reproducible phenotyping |
| title_full | A closer look at Aspergillus: online monitoring via scattered light enables reproducible phenotyping |
| title_fullStr | A closer look at Aspergillus: online monitoring via scattered light enables reproducible phenotyping |
| title_full_unstemmed | A closer look at Aspergillus: online monitoring via scattered light enables reproducible phenotyping |
| title_short | A closer look at Aspergillus: online monitoring via scattered light enables reproducible phenotyping |
| title_sort | closer look at aspergillus online monitoring via scattered light enables reproducible phenotyping |
| topic | Aspergillus giganteus Microtiter plate Miniaturized cultivation Antifungal protein High throughput bioprocess development Morphology engineering |
| url | http://link.springer.com/article/10.1186/s40694-019-0073-x |
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